背景:弓形虫感染可导致严重的肺炎,目前的治疗方法非常不充分。NLRP3炎性体是具有pyrin结构域的NOD样受体家族的成员之一,这对弓形虫的先天免疫防御至关重要。研究表明,白藜芦醇(RSV)通过抑制弓形虫来源的热休克蛋白70/TLR4/NF-κB途径来预防由这种感染引起的肺损伤,从而减少巨噬细胞驱动的炎症反应。然而,应该提到的是,NLRP3炎性体参与弓形虫感染引起的肺损伤的免疫反应尚不完全清楚。
目的:本研究旨在阐明RSV如何改善弓形虫感染引起的肺损伤,特别关注涉及TLR4,NF-κB的途径,和NLRP3炎性体。
方法:通过在BALB/c小鼠中使用弓形虫的RH菌株和RAW264.7巨噬细胞系来建立体外和体内感染模型。利用分子对接等技术探讨RSV的作用机制,表面等离子体共振,ELISA,蛋白质印迹,免疫共沉淀,和免疫荧光染色。
结果:研究结果表明,TLR4或NF-κB的抑制会影响与NLRP3炎性体途径相关的蛋白质水平。此外,观察到对RSV和NLRP3之间的结合的显著亲和力。RSV治疗导致肺组织和RAW264.7细胞内NLRP3炎性体的激活和形成显着减少,同时支气管肺泡灌洗液中IL-1β浓度降低。这些结果与使用NLRP3抑制剂CY-09时观察到的结果一致。此外,在RSV之前应用CY-09否定了后者的抗炎特性。
结论:考虑到先前研究的见解以及当前研究的结果,TLR4/NF-κB/NLRP3信号通路似乎成为免疫调节减轻弓形虫感染引起的肺损伤的有希望的靶标。本研究中收集的证据为RSV作为具有抗弓形虫特性和调节炎症反应能力的治疗剂的持续探索和潜在的未来临床部署奠定了基础。
BACKGROUND: Infection by
Toxoplasma gondii can lead to severe pneumonia, with current treatments being highly inadequate. The NLRP3 inflammasome is one member of the NOD-like receptor family with a pyrin domain, which is crucial in the innate immune defense against T. gondii. Research has shown that resveratrol (RSV) prevents lung damage caused by this infection by inhibiting the T. gondii-derived heat shock protein 70/TLR4/NF-κB pathway, thus reducing the macrophage-driven inflammatory response. However, it should be mentioned that the participation of NLRP3 inflammasome in the immune response to the lung injuries caused by T. gondii infections is not entirely clear.
OBJECTIVE: This study aims to clarify how RSV ameliorates lung damage triggered by
Toxoplasma gondii infection, with a particular focus on the pathway involving TLR4, NF-κB, and the NLRP3 inflammasome.
METHODS: Both in vitro and in vivo models of infection were developed by employing the RH strain of T. gondii in BALB/c mice and RAW 264.7 macrophage cell lines. The action mechanism of RSV was explored using techniques such as molecular docking, surface plasmon resonance, ELISA, Western blot, co-immunoprecipitation, and immunofluorescence staining.
RESULTS: Findings indicate that the suppression of TLR4 or NF-κB impacts the levels of proteins associated with the NLRP3 inflammasome pathway. Additionally, a significant affinity for binding between RSV and NLRP3 was observed. Treatment with RSV led to a marked reduction in the activation and formation of the NLRP3 inflammasome within lung tissues and RAW 264.7 cells, alongside a decrease in IL-1β concentrations in the bronchoalveolar lavage fluid. These outcomes align with those seen when using the NLRP3 inhibitor CY-09. Moreover, the application of CY-09 prior to RSV negated the latter\'s anti-inflammatory properties.
CONCLUSIONS: Considering insights from previous research alongside the outcomes of the current investigation, it appears that the TLR4/NF-κB/NLRP3 signaling pathway emerges as a promising target for immunomodulation to alleviate lung injury from T. gondii infection. The evidence gathered in this study lays the groundwork for the continued exploration and potential future clinical deployment of RSV as a therapeutic agent with anti-
Toxoplasma properties and the capability to modulate the inflammatory response.